期刊论文详细信息
BMC Genomics
Unusual genome complexity in Lactobacillus salivarius JCM1046
Paul W O’Toole1  Marcus J Claesson1  Brian M Forde1  Emma J Raftis1 
[1] Alimentary Pharmabiotic Centre, University College Cork, Cork, Ireland
关键词: Conjugative transposon;    Tn6224;    Linear plasmid;    Multireplicon;    Megaplasmid;    Lactobacillus salivarius;   
Others  :  1140801
DOI  :  10.1186/1471-2164-15-771
 received in 2014-05-21, accepted in 2014-08-26,  发布年份 2014
PDF
【 摘 要 】

Background

Lactobacillus salivarius strains are increasingly being exploited for their probiotic properties in humans and animals. Dissemination of antibiotic resistance genes among species with food or probiotic-association is undesirable and is often mediated by plasmids or integrative and conjugative elements. L. salivarius strains typically have multireplicon genomes including circular megaplasmids that encode strain-specific traits for intestinal survival and probiotic activity. Linear plasmids are less common in lactobacilli and show a very limited distribution in L. salivarius. Here we present experimental evidence that supports an unusually complex multireplicon genome structure in the porcine isolate L. salivarius JCM1046.

Results

JCM1046 harbours a 1.83 Mb chromosome, and four plasmids which constitute 20% of the genome. In addition to the known 219 kb repA-type megaplasmid pMP1046A, we identified and experimentally validated the topology of three additional replicons, the circular pMP1046B (129 kb), a linear plasmid pLMP1046 (101 kb) and pCTN1046 (33 kb) harbouring a conjugative transposon. pMP1046B harbours both plasmid-associated replication genes and paralogues of chromosomally encoded housekeeping and information-processing related genes, thus qualifying it as a putative chromid. pLMP1046 shares limited sequence homology or gene synteny with other L. salivarius plasmids, and its putative replication-associated protein is homologous to the RepA/E proteins found in the large circular megaplasmids of L. salivarius. Plasmid pCTN1046 harbours a single copy of an integrated conjugative transposon (Tn6224) which appears to be functionally intact and includes the tetracycline resistance gene tetM.

Conclusion

Experimental validation of sequence assemblies and plasmid topology resolved the complex genome architecture of L. salivarius JCM1046. A high-coverage draft genome sequence would not have elucidated the genome complexity in this strain. Given the expanding use of L. salivarius as a probiotic, it is important to determine the genotypic and phenotypic organization of L. salivarius strains. The identification of Tn6224-like elements in this species has implications for strain selection for probiotic applications.

【 授权许可】

   
2014 Raftis et al.; licensee BioMed Central Ltd.

【 预 览 】
附件列表
Files Size Format View
20150325113236425.pdf 3008KB PDF download
Figure 4. 78KB Image download
Figure 3. 165KB Image download
Figure 2. 98KB Image download
Figure 1. 111KB Image download
【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

【 参考文献 】
  • [1]Li Y, Raftis E, Canchaya C, Fitzgerald GF, Sinderen DV, O’Toole PW: Polyphasic analysis indicates that Lactobacillus salivarius subsp. salivarius and Lactobacillus salivarius subsp. salicinius do not merit separate subspecies status. Int J of Syst Evol Microbiol 2006, 56:2397-2403.
  • [2]Reuter G: The Lactobacillus and Bifidobacterium microflora of the human intestine: composition and succession. Curr Issues Intest Microbiol 2001, 2(2):43-53.
  • [3]Mitsuoka T: Vergleichende untersuchungen Über die Laktobazillen aus den faeces von menschen, schweinen und hÜhnern. Bakteriol 1969, 210:32-51.
  • [4]Martín R, Jiménez E, Olivares M, Marín ML, Fernández L, Xaus J, Rodríguez JM: Lactobacillus salivarius CECT 5713, a potential probiotic strain isolated from infant feces and breast milk of a mother-child pair. Int J Food Microbiol 2006, 112(1):35-43.
  • [5]Neville BA, O'Toole PW: Probiotic properties of Lactobacillus salivarius and closely related Lactobacillus species. Future Microbiol 2010, 5(5):759-774.
  • [6]Ryan KA, Daly P, Li Y, Hooton C, O'Toole PW: Strain-specific inhibition of Helicobacter pylori by Lactobacillus salivarius and other lactobacilli. J Antimicrob Chemother 2008, 61(4):831-834.
  • [7]Raftis EJ, Salvetti E, Torriani S, Felis GE, O'Toole PW: Genomic diversity of Lactobacillus salivarius. Appl Environ Microbiol 2011, 77(3):954-965.
  • [8]Flynn S, van Sinderen D, Thornton GM, Holo H, Nes IF, Collins JK: Characterization of the genetic locus responsible for the production of ABP-118, a novel bacteriocin produced by the probiotic bacterium Lactobacillus salivarius subsp. salivarius UCC118. Microbiology 2002, 148(4):973-984.
  • [9]Claesson MJ, Li Y, Leahy S, Canchaya C, van Pijkeren JP, Cerdeño-Tárraga AM, Parkhill J, Flynn S, O’Sullivan GC, Collins JK, Higgins D, Shanahan F, Fitzgerald GF, van Sinderen D, O'Toole PW: Multireplicon genome architecture of Lactobacillus salivarius. Proc Natl Acad Sci 2006, 103(17):6718-6723.
  • [10]Corr SC, Li Y, Riedel CU, O'Toole PW, Hill C, Gahan CGM: Bacteriocin production as a mechanism for the antiinfective activity of Lactobacillus salivarius UCC118. Proc Natl Acad Sci 2007, 104(18):7617-7621.
  • [11]Fang F, Li Y, Bumann M, Raftis EJ, Casey PG, Cooney JC, Walsh MA, O'Toole PW: Allelic variation of bile salt hydrolase genes in Lactobacillus salivarius does not determine bile resistance levels. J Bact 2009, 191(18):5743-5757.
  • [12]Li Y, Canchaya C, Fang F, Raftis E, Ryan KA, van Pijkeren J-P, van Sinderen D, O'Toole PW: Distribution of megaplasmids in Lactobacillus salivarius and other lactobacilli. J Bacteriol 2007, 189(17):6128-6139.
  • [13]van Pijkeren J, Canchaya C, Ryan K, Li Y, Claesson M, Sheil B, Steidler L, O'Mahony L, Fitzgerald G, van Sinderen D: Comparative and functional analysis of sortase-dependent proteins in the predicted secretome of Lactobacillus salivarius UCC118. Appl Environ Microbiol 2006, 72(6):4143-4153.
  • [14]Jimenez E, Martin R, Maldonado A, Martin V, Gomez de Segura A, Fernandez L, Rodriguez JM: Complete genome sequence of Lactobacillus salivarius CECT 5713, a probiotic strain isolated from human milk and infant feces. J Bacteriol 2010, 192(19):5266-5267.
  • [15]Ham J-S, Kim H-W, Seol K-H, Jang A, Jeong S-G, Oh M-H, Kim D-H, Kang D-K, Kim G-B, Cha C-J: Genome sequence of Lactobacillus salivarius NIAS840, isolated from chicken intestine. J Bacteriol 2011, 193(19):5551-5552.
  • [16]Kergourlay G, Messaoudi S, Dousset X, Prévost H: Genome Sequence of Lactobacillus salivarius SMXD51, a Potential Probiotic Strain Isolated from Chicken Cecum, Showing Anti-Campylobacter Activity. J Bacteriol 2012, 194(11):3008-3009.
  • [17]Cho Y-J, Choi JK, Kim J-H, Lim Y-S, Ham J-S, Kang D-K, Chun J, Paik H-D, Kim G-B: Genome sequence of Lactobacillus salivarius GJ-24, a probiotic strain isolated from healthy adult intestine. J Bacteriol 2011, 193(18):5021-5022.
  • [18]MacKenzie DA, McLay K, Roos S, Walter J, Swarbreck D, Drou N, Crossman LC, Juge N: Draft Genome Sequence of a Novel Lactobacillus salivarius Strain Isolated from Piglet. Genome Announcements 2014, 2:1.
  • [19]Overhage J, Sielker S, Homburg S, Parschat K, Fetzner S: Identification of large linear plasmids in Arthrobacter spp. encoding the degradation of quinaldine to anthranilate. Microbiology 2005, 151(2):491-500.
  • [20]Kinashi H: Giant linear plasmids in Streptomyces: a treasure trove of antibiotic biosynthetic clusters. J Antibiot 2011, 64(1):19-25.
  • [21]Chater K, Kinashi H: Streptomyces; linear plasmids: their discovery, functions, interactions with other replicons, and evolutionary significance. In Microbial Linear Plasmids, Volume 7. Edited by Meinhardt F, Klassen R. Heidelberg: Springer Berlin; 2007:1-31.
  • [22]Chen C: Streptomyces; linear plasmids: replication and telomeres. In Microbial Linear Plasmids, Volume 7. Edited by Meinhardt F, Klassen R. Springer Berlin: Heidelberg; 2007:33-61.
  • [23]Barbour A, Garon C: Linear plasmids of the bacterium Borrelia burgdorferi have covalently closed ends. Science 1987, 237(4813):409-411.
  • [24]Stromsten NJ, Benson SD, Burnett RM, Bamford DH, Bamford JKH: The Bacillus thuringiensis linear double-stranded DNA phage Bam35, which Is highly similar to the Bacillus cereus linear plasmid pBClin15, has a prophage state. J Bacteriol 2003, 185(23):6985-6989.
  • [25]Ravin NV: N15: The linear phage-plasmid. Plasmid 2011, 65(2):102-109.
  • [26]Hertwig S, Klein I, Lurz R, Lanka E, Appel B: PY54, a linear plasmid prophage of Yersinia enterocolitica with covalently closed ends. Mol Microbiol 2003, 48(4):989-1003.
  • [27]Casjens SR, Gilcrease EB, Huang WM, Bunny KL, Pedulla ML, Ford ME, Houtz JM, Hatfull GF, Hendrix RW: The pKO2 linear plasmid prophage of Klebsiella oxytoca. J Bacteriol 2004, 186(6):1818-1832.
  • [28]Alemayehu D, Ross RP, O'Sullivan O, Coffey A, Stanton C, Fitzgerald GF, McAuliffe O: Genome of a virulent bacteriophage Lb338-1 that lyses the probiotic Lactobacillus paracasei cheese strain. Gene 2009, 448(1):29-39.
  • [29]Roussel Y, Colmin C, Simonet JM, Decaris B: Strain characterization, genome size and plasmid content in the Lactobacillus acidophilus group (Hansen and Mocquot). J Appl Bacteriol 1993, 74:549-556.
  • [30]Franke AE, Clewell DB: Evidence for a chromosome-borne resistance transposon (Tn916) in Streptococcus faecalis that is capable of "conjugal" transfer in the absence of a conjugative plasmid. J Bacteriol 1981, 145(1):494-502.
  • [31]Clewell DB, Flannagan SE, Jaworski DD: Unconstrained bacterial promiscuity: the Tn916-Tn1545 family of conjugative transposons. Trends Microbiol 1995, 3(6):229-236.
  • [32]Bertram J, Stratz M, Durre P: Natural transfer of conjugative transposon Tn916 between gram-positive and gram-negative bacteria. J Bacteriol 1991, 173(2):443-448.
  • [33]Boguslawska J, Zycka-Krzesinska J, Wilcks A, Bardowski J: Intra- and interspecies conjugal transfer of Tn916-like elements from Lactococcus lactis in vitro and in vivo. Appl Environ Microbiol 2009, 75(19):6352-6360.
  • [34]Devirgiliis C, Coppola D, Barile S, Colonna B, Perozzi G: Characterization of the Tn916 conjugative transposon in a food-borne strain of Lactobacillus paracasei. Appl Environ Microbiol 2009, 75(12):3866-3871.
  • [35]Roberts AP, Cheah G, Ready D, Pratten J, Wilson M, Mullany P: Transfer of Tn916-like elements in microcosm dental plaques. Antimicrob Agents Chemother 2001, 45(10):2943-2946.
  • [36]Schjørring S, Krogfelt KA: Assessment of bacterial antibiotic resistance transfer in the gut. Int J Microbiol 2010, 2011:Article ID 312956.
  • [37]Wozniak R, Waldor M: Integrative and conjugative elements: mosaic mobile genetic elements enabling dynamic lateral gene flow. Nature reviews Microbiology 2010, 8(8):552-563.
  • [38]Roberts AP, Mullany P: A modular master on the move: the Tn916 family of mobile genetic elements. Trends Microbiol 2009, 17(6):251-258.
  • [39]Carver T, Thomson N, Bleasby A, Berriman M, Parkhill J: DNAPlotter: circular and linear interactive genome visualization. Bioinformatics 2009, 25:119-120.
  • [40]Fang F, Flynn S, Li Y, Claesson MJ, van Pijkeren J-P, Collins JK, van Sinderen D, O'Toole PW: Characterization of endogenous plasmids from Lactobacillus salivarius UCC118. Appl Environ Microbiol 2008, 74(10):3216-3228.
  • [41]Barton BM, Harding GP, Zuccarelli AJ: A general method for detecting and sizing large plasmids. Analyst Biochem 1995, 226(2):235-240.
  • [42]Warner JE, Onderdonk AB: Method for Optimizing Pulsed-Field Gel Electrophoresis Banding Pattern Data. JMD 2003, 5(1):21-27.
  • [43]Wang H, Roberts AP, Mullany P: DNA sequence of the insertional hot spot of Tn916 in the Clostridium difficile genome and discovery of a Tn916-like element in an environmental isolate integrated in the same hot spot. FEMS Microbiol Lett 2000, 192(1):15-20.
  • [44]Harrison PW, Lower RPJ, Kim NKD, Young JPW: Introducing the bacterial chromid”: not a chromosome, not a plasmid. Trends Microbiol 2010, 18(4):141-148.
  • [45]Sanchez-Perez G, Mira A, Nyirő G, Pašić L, Rodriguez-Valera F: Adapting to environmental changes using specialized paralogs. Trends Genet 2008, 24(4):154-158.
  • [46]Wagenknecht M, Dib J, Thürmer A, Daniel R, Farías M, Meinhardt F: Structural peculiarities of linear megaplasmid, pLMA1, from Micrococcus luteus; interfere with pyrosequencing reads assembly. Biotechnol Lett 2010, 32(12):1853-1862.
  • [47]Roberts AP, Chandler M, Courvalin P, Guédon G, Mullany P, Pembroke T, Rood JI, Jeffery Smith C, Summers AO, Tsuda M, Berg DE: Revised nomenclature for transposable genetic elements. Plasmid 2008, 60(3):167-173.
  • [48]Zhou Y, Liang Y, Lynch KH, Dennis JJ, Wishart DS: PHAST: A fast phage search tool. Nucleic Acids Res 2011, 39(suppl 2):W347-W352.
  • [49]Jore MM, Brouns SJJ, van der Oost J: RNA in Defense: CRISPRs Protect Prokaryotes against Mobile Genetic Elements. Cold Spring Harb Perspect Biol 2012., 4(6) http://cshperspectives.cshlp.org/content/4/6/a003657.full.pdf+html webcite
  • [50]Jannière L, Gruss A, Ehrlich D: Plasmids. In Bacillus subtilis and other gram-positive bacteria. Edited by Sonenshein JAH AL, Losick R. Washington, D.C: American Society for Microbiology; 1993:625-644.
  • [51]Alikhan N-F, Petty N, Ben Zakour N, Beatson S: BLAST Ring Image Generator (BRIG): simple prokaryote genome comparisons. BMC Genomics 2011, 12(1):402. BioMed Central Full Text
  • [52]Bentley SD, Parkhill J: Comparative genomic structure of prokaryotes. Annu Rev Genet 2004, 38(1):771-791.
  • [53]Heavens D, Tailford LE, Crossman L, Jeffers F, MacKenzie DA, Caccamo M, Juge N: Genome Sequence of the Vertebrate Gut Symbiont Lactobacillus reuteri ATCC 53608. J Bacteriol 2011, 193(15):4015-4016.
  • [54]Beaurepaire C, Chaconas G: Mapping of essential replication functions of the linear plasmid lp17 of B. burgdorferi by targeted deletion walking. Mol Microbiol 2005, 57(1):132-142.
  • [55]Chang P-C, Cohen SN: Birdirectional replication from an internal origin in a linear Streptomyces plasmid. Science 1994, 265:952-954.
  • [56]Ravin NV, Kuprianov VV, Gilcrease EB, Casjens SR: Bidirectional replication from an internal ori site of the linear N15 plasmid prophage. Nucleic Acids Res 2003, 31(22):6552-6560.
  • [57]Mardanov A, Ravin N: Functional characterization of the repA replication gene of linear plasmid prophage N15. Res Microbiol 2006, 157(2):176-183.
  • [58]Shiffman D, Cohen SN: Reconstruction of Streptomyces linear plasmid replication from separately cloned DNA fragment: existence pf a cryptic origin of circular replication within the linear plasmid. Proc Natl Acad Sci U S A 1992, 89(89):6129-6133.
  • [59]Laing C, Buchanan C, Taboada E, Zhang Y, Kropinski A, Villegas A, Thomas J, Gannon V: Pan-genome sequence analysis using Panseq: an online tool for the rapid analysis of core and accessory genomic regions. BMC Bioinformatics 2010, 11(1):461. BioMed Central Full Text
  • [60]Berger B, Pridmore R, Barretto C, Delmas-Julien F, Schreiber K, Arigoni F, Brüssow H: Similarity and Differences in the Lactobacillus acidophilus Group Identified by Polyphasic Analysis and Comparative Genomics. J Bacteriol 2007, 189(4):1311-1321.
  • [61]Joyce SA, MacSharry J, Casey PG, Kinsella M, Murphy EF, Shanahan F, Hill C, Gahan CGM: Regulation of host weight gain and lipid metabolism by bacterial bile acid modification in the gut. Proc Natl Acad Sci 2014, 111(20):7421-7426.
  • [62]Messaoudi S, Manai M, Kergourlay G, Prévost H, Connil N, Chobert JM, Dousset X: Lactobacillus salivarius: Bacteriocin and probiotic activity. Food Microbiol 2013, 36(2):296-304.
  • [63]Barrett E, Hayes M, O'Connor P, Gardiner G, Fitzgerald GF, Stanton C, Ross RP, Hill C: Salivaricin P, one of a family of two-component antilisterial bacteriocins produced by intestinal isolates of Lactobacillus salivarius. Appl Environ Microbiol 2007, 73(11):3719-3723.
  • [64]O'Shea EF, O'Connor PM, Raftis EJ, O'Toole PW, Stanton C, Cotter PD, Ross RP, Hill C: Production of Multiple Bacteriocins from a Single Locus by Gastrointestinal Strains of Lactobacillus salivarius. J Bacteriol 2011, 193(24):6973-6982.
  • [65]Chen K, Reuter M, Sanghvi B, Roberts GA, Cooper LP, Tilling M, Blakely GW, Dryden DTF: ArdA proteins from different mobile genetic elements can bind to the EcoKI Type I DNA methyltransferase of E. coli K12. Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics 2014, 1844(3):505-511.
  • [66]Guérillot R, Da Cunha V, Sauvage E, Bouchier C, Glaser P: Modular evolution of TnGBSs, a new family of integrative and conjugative elements associating insertion sequence transposition, plasmid replication, and conjugation for their spreading. J Bacteriol 2013, 195(9):1979-1990.
  • [67]Panel F: Opinion of the Scientific Panel on additives and products or substances used in animal feed (FEEDAP) on the updating of the criteria used in the assessment of bacteria for resistance to antibiotics of human or veterinary importance. EFSA J 2005, 223:1-12.
  • [68]Zerbino DR, Birney E: Velvet: Algorithms for de novo short read assembly using de Bruijn graphs. Genome Res 2008, 18(5):821-829.
  • [69]Mardis ER: Next-Generation DNA Sequencing Methods. Annu Rev Genomics Hum Genet 2008, 9(1):387-402.
  • [70]Chevreux B, Wetter TSS: Genome sequence assembly using trace signals and additional sequence information. Computer Science and Biology: Proceedings of the German Conference on Bioinformatics (GCB) 1999, 99:45-56.
  • [71]Green P: PHRAP v 1.080812. 1999. [http://phrap.org webcite]
  • [72]Milne I, Bayer M, Cardle L, Shaw P, Stephen G, Wright F, Marshall D: Tablet—next generation sequence assembly visualization. Bioinformatics 2010, 26(3):401-402.
  • [73]Forde B, Neville B, O' Donnell M, Riboulet-Bisson E, Claesson M, Coghlan A, Ross R, O' Toole P: Genome sequences and comparative genomics of two Lactobacillus ruminis strains from the bovine and human intestinal tracts. Microb Cell Fact 2011, 10(1):S13. BioMed Central Full Text
  • [74]Otto TD, Dillon GP, Degrave WS, Berriman M: RATT: Rapid Annotation Transfer Tool. Nucleic Acids Res 2011.
  • [75]Rutherford K, Parkhill J, Crook J, Horsnell T, Rice P, Rajandream M-A, Barrell B: Artemis: sequence visualization and annotation. Bioinformatics 2000, 16(10):944-945.
  • [76]Carver TJ, Rutherford KM, Berriman M, Rajandream MA, Barrell BG, Parkhill J: ACT: the Artemis comparison tool. Bioinformatics 2005, 21:3422-3423.
  文献评价指标  
  下载次数:27次 浏览次数:26次